BRENDA - Enzyme Database show

Kinetic analysis and ligand-induced conformational changes in dimeric and tetrameric forms of human thymidine kinase 2

Barroso, J.F.; Carvalho, R.N.; Flatmark, T.; Biochemistry 44, 4886-4896 (2005)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.21
DNA and amino acid sequence determination, expression in Escherichia coli as maltose binding fusion protein in inclusion bodies, coexpression of GroEL/ES molecular chaperone system and 3% v/v ethanol supplementation
Homo sapiens
General Stability
EC Number
General Stability
Organism
2.7.1.21
binding of dNTP inhibitors and substrates, i.e. 2'-deoxycytidine, Mg-2'-deoxyCTP, 2'-deoxythymidine, and Mg-2'-deoxyTTP structurally stabilizes the enzyme, ligand-bound enzyme is less susceptible to aggregation, proteolysis by trypsin, and thermal denaturation
Homo sapiens
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.21
dATP
strong feedback inhibition, tight binding at the active site in stoichiometric amount
Homo sapiens
2.7.1.21
dCTP
strong feedback inhibition, tight binding at the active site in stoichiometric amount
Homo sapiens
2.7.1.21
dTTP
strong feedback inhibition, tight binding at the active site in stoichiometric amount
Homo sapiens
2.7.1.21
additional information
inhibition potency of dNTPs in descending order: dTTP, dCTP, dATP, inhibitor binding structurally stabilizes the enzyme, induction of conformational changes upon ligand binding, overview
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
additional information
-
additional information
steady-state kinetics, kinetics of conformational changes induced by ligand binding
Homo sapiens
2.7.1.21
0.0028
-
MgATP2-
dNTP-free recombinant dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.004
-
2'-deoxycytidine
dNTP-free recombinant dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0049
-
2'-deoxycytidine
dNTP-free recombinant tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0058
-
2'-deoxythymidine
dNTP-free recombinant dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0059
-
MgATP2-
dNTP-free recombinant tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0065
-
2'-deoxythymidine
dNTP-free recombinant tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.21
Mg2+
-
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
Homo sapiens
-
ADP + 2'-deoxycytidine 5'-phosphate
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
Homo sapiens
-
ADP + 2'-deoxythymidine 5'-phosphate
-
-
ir
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.21
Homo sapiens
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.21
recombinant enzyme as maltose binding fusion protein from Escherichia coli
Homo sapiens
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.21
additional information
-
-
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
-
661195
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxycytidine
induction of conformational changes upon ligand binding, overview
661195
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
-
661195
Homo sapiens
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
induction of conformational changes upon ligand binding, overview
661195
Homo sapiens
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
ir
2.7.1.21
additional information
ligand-induced conformational changes in dimeric and tetrameric forms of the enzyme alter the catalytic activity
661195
Homo sapiens
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
2.7.1.21
dimer
the most abundant enzyme form, less active than the tetramer
Homo sapiens
2.7.1.21
More
ligand-induced conformational changes in dimeric and tetrameric forms of the enzyme
Homo sapiens
2.7.1.21
tetramer
less abundant form, more active than the dimer
Homo sapiens
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.21
25
-
assay at
Homo sapiens
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.1.21
additional information
-
irreversible thermal denaturation following a two-state irrversible denaturation model, study with ligand-bound and free enzyme, respectively, overview
Homo sapiens
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.21
7.5
-
assay at
Homo sapiens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.21
MgATP2-
-
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
DNA and amino acid sequence determination, expression in Escherichia coli as maltose binding fusion protein in inclusion bodies, coexpression of GroEL/ES molecular chaperone system and 3% v/v ethanol supplementation
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.21
MgATP2-
-
Homo sapiens
General Stability (protein specific)
EC Number
General Stability
Organism
2.7.1.21
binding of dNTP inhibitors and substrates, i.e. 2'-deoxycytidine, Mg-2'-deoxyCTP, 2'-deoxythymidine, and Mg-2'-deoxyTTP structurally stabilizes the enzyme, ligand-bound enzyme is less susceptible to aggregation, proteolysis by trypsin, and thermal denaturation
Homo sapiens
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.21
dATP
strong feedback inhibition, tight binding at the active site in stoichiometric amount
Homo sapiens
2.7.1.21
dCTP
strong feedback inhibition, tight binding at the active site in stoichiometric amount
Homo sapiens
2.7.1.21
dTTP
strong feedback inhibition, tight binding at the active site in stoichiometric amount
Homo sapiens
2.7.1.21
additional information
inhibition potency of dNTPs in descending order: dTTP, dCTP, dATP, inhibitor binding structurally stabilizes the enzyme, induction of conformational changes upon ligand binding, overview
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
additional information
-
additional information
steady-state kinetics, kinetics of conformational changes induced by ligand binding
Homo sapiens
2.7.1.21
0.0028
-
MgATP2-
dNTP-free recombinant dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.004
-
2'-deoxycytidine
dNTP-free recombinant dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0049
-
2'-deoxycytidine
dNTP-free recombinant tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0058
-
2'-deoxythymidine
dNTP-free recombinant dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0059
-
MgATP2-
dNTP-free recombinant tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0065
-
2'-deoxythymidine
dNTP-free recombinant tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.21
Mg2+
-
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
Homo sapiens
-
ADP + 2'-deoxycytidine 5'-phosphate
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
Homo sapiens
-
ADP + 2'-deoxythymidine 5'-phosphate
-
-
ir
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
recombinant enzyme as maltose binding fusion protein from Escherichia coli
Homo sapiens
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.21
additional information
-
-
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
-
661195
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxycytidine
induction of conformational changes upon ligand binding, overview
661195
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
-
661195
Homo sapiens
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
induction of conformational changes upon ligand binding, overview
661195
Homo sapiens
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
ir
2.7.1.21
additional information
ligand-induced conformational changes in dimeric and tetrameric forms of the enzyme alter the catalytic activity
661195
Homo sapiens
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.1.21
dimer
the most abundant enzyme form, less active than the tetramer
Homo sapiens
2.7.1.21
More
ligand-induced conformational changes in dimeric and tetrameric forms of the enzyme
Homo sapiens
2.7.1.21
tetramer
less abundant form, more active than the dimer
Homo sapiens
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.21
25
-
assay at
Homo sapiens
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.1.21
additional information
-
irreversible thermal denaturation following a two-state irrversible denaturation model, study with ligand-bound and free enzyme, respectively, overview
Homo sapiens
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.21
7.5
-
assay at
Homo sapiens